I was a bit doubtful about the hype (may make directly labeled mAbs obsolete), but a week after seeing Richard Haugland's posting about Zenon Fab labeling kits I suddenly faced a situation perfectly suited for their use. We were faced with 4 steps staining protocol, but the Zenon reagent reduced this to one step. Basically these are goat anti-mouse Fab fragments that allow quick 10 minute labeling of primary mAbs; a picture is worth a thousand words (http://www.probes.com/products/zenon/zenon.html). We have been using the Zenon APC reagent for intracellular staining of eosinophils for the past 3 weeks and have been getting very good results with low non-specific binding and bright signal. The Alexa 647 seems to bind eosinophils non-specifically, but this is likely due to the eosinophil, as FITC also binds eos. Many cutting edge mAbs are simply not commercially available as flurochrome conjugates and certainly not as exotic fluorochrome conjugates. This systems allows one a lot more flexibility in experimental design. Not meaning to be overly promotional, but I am interested in other's experiences with or thoughts about these reagents and their strengths and limitations. I'm sure many folks out there could benefit from such a discussion. Calman > _______________________ > Calman Prussin > Laboratory of Allergic Diseases > NIAID/ National Institutes of Health >
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